[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

EP1930025A1 - VEHICULE POUR ANTIGENES ET MEDICAMENTS PERMETTANT DE BASCULER DE LA PRODUCTION SELECTIVE D'ANTICORPS IgA A LA PRODUCTION D' ANTICORPS TANT IgA QUE IgG, ET VACCIN TRANSNASAL/TRANSMUCOSIQUE UTILISANT LE VEHICULE - Google Patents

VEHICULE POUR ANTIGENES ET MEDICAMENTS PERMETTANT DE BASCULER DE LA PRODUCTION SELECTIVE D'ANTICORPS IgA A LA PRODUCTION D' ANTICORPS TANT IgA QUE IgG, ET VACCIN TRANSNASAL/TRANSMUCOSIQUE UTILISANT LE VEHICULE Download PDF

Info

Publication number
EP1930025A1
EP1930025A1 EP06782373A EP06782373A EP1930025A1 EP 1930025 A1 EP1930025 A1 EP 1930025A1 EP 06782373 A EP06782373 A EP 06782373A EP 06782373 A EP06782373 A EP 06782373A EP 1930025 A1 EP1930025 A1 EP 1930025A1
Authority
EP
European Patent Office
Prior art keywords
vehicle
vaccine
antigen
iga
seq
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP06782373A
Other languages
German (de)
English (en)
Other versions
EP1930025B1 (fr
EP1930025A4 (fr
Inventor
Hiroshi Kido
Dai Mizuno
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Applied Medical Enzyme Research Institute Corporat
University of Tokushima NUC
Original Assignee
University of Tokushima NUC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Tokushima NUC filed Critical University of Tokushima NUC
Publication of EP1930025A1 publication Critical patent/EP1930025A1/fr
Publication of EP1930025A4 publication Critical patent/EP1930025A4/fr
Application granted granted Critical
Publication of EP1930025B1 publication Critical patent/EP1930025B1/fr
Ceased legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/145—Orthomyxoviridae, e.g. influenza virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/35—Allergens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/385—Haptens or antigens, bound to carriers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5252—Virus inactivated (killed)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • A61K2039/541—Mucosal route
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • A61K2039/541—Mucosal route
    • A61K2039/542—Mucosal route oral/gastrointestinal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • A61K2039/541—Mucosal route
    • A61K2039/543—Mucosal route intranasal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55516—Proteins; Peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55555—Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • This invention relates to an antigen-drug (AD) vehicle enabling transnasal, transmucosal, and transdermal administration and a method for inducting mucosal immunity and humoral immunity using the AD vehicle, more specifically, to an AD vehicle enabling a switch from induction of selective production of IgA antibody to induction of production of both of IgA and IgG antibodies and a transnasal vaccine, a mucosal vaccine, a therapeutic/prophylactic agent of allergy, and the like using the AD vehicle.
  • AD antigen-drug
  • the conventional inactivated vaccines, toxoids, and the like have the function and effect of enhancing humoral immunity mainly by inducing production of IgG antibody in the blood of vaccinated subjects, and the efficacy thereof has been confirmed.
  • the conventional inactivated vaccines and toxoids have the low IgA antibody production and low mucosal immunity-inducing capacity, they have a limit from the viewpoints of satisfactory function and effect for preventing natural infection. Under the circumstances, many attempts have been made in various aspects for solving the drawbacks of conventional vaccines.
  • the necessity of a switch from the conventional vaccine to be inoculated subcutaneously, intramuscularly, or the like to the mucosal vaccine inducing production of IgA antibody at the mucosae which are the natural virus infection routes is widely and deeply recognized.
  • mucosal vaccine as a new generation vaccine in 21st century for inducing production of IgA antibody and local immunity or mucosal immunity are expected worldwide, such mucosal vaccine has not been achieved. It is considered that the mucosal vaccine has not been achieved since a safe and effective adjuvant for imparting the function of inducing IgA antibody production and local immunity or mucosal immunity has not been specified nor established.
  • Patent Document 1 JP-T-2002-521460
  • Patent Document 2 JP-A-3-135923
  • Patent Document 3 JP-T-10-500102
  • Patent Document 4 JP-T-2001-523729
  • Patent Document 5 JP-T-2002-50452
  • Patent Document 6 JP-A-2003-116385
  • Patent Document 7 JP-A-2003-50452
  • Patent Document 8 PCT WO00/20039 pamphlet
  • Non-Patent Document 1 New Engl. J. Med., Vol. 350, pages 896-903, 2004
  • Non-Patent Document 2 New Engl. J. Med., Vol. 350, pages 860-861, 2004
  • Non-Patent Document 3 Cleve. Clin. J. Med., Vol.
  • Non-Patent Document 4 Am. J. Respir. Cell Mol. Biol., Vol. 24, 452-458, 2001
  • Non-Patent Document 5 AdV/Ances Drug Delivery Rev., Vol. 51, pages 81-96, 2001
  • Non-Patent Document 6 V/Accine, Vol. 21, 3706-3712, 2003
  • Non-Patent Document 7 Infection and Immunity, Vol. 71, pages 4780-4788, 2003
  • Non-Patent Document 8 J. neonatal Nursing, Vol. 10, pages 2-11, 2004
  • Non-Patent Document 9 Biology of the Neonate, Vol. 74 (suppl1), pages 9-14, 1998
  • This invention provides an AD vehicle enabling transnasal, transmucosal, and transdermal administrations, a method for inducing mucosal immunity and humoral immunity using the AD vehicle, more specifically, an AD vehicle enabling a switch from induction of selective production of IgA antibody to induction of production of both of IgA and IgG antibodies and a transnasal vaccine, a mucosal vaccine, a therapeutic/prophylactic agent for allergy, and the like using the AD vehicle.
  • AD vehicle to be provided by this invention enable realization and diffusion of a mucosal vaccine, a prophylactic/therapeutic agent for allergy, and transmucosal/transdermal drug administration.
  • the transnasal/mucosal vaccine exhibits a considerably excellent infection prevention effect as compared to conventional vaccines since it is an immunization means adapted to the actual condition of natural infection.
  • mucosal IgA and IgG in nostrils induced by the AD vehicle causes inactivation of an allergen at the site to enable hyposensitization.
  • application of the AD vehicle to various drugs reinforces and promotes prophylactic/therapeutic effects by transmucosal administration and transdermal administration of the drugs.
  • this invention greatly improves medical practice/healthcare/hygiene of all humankind and is much-anticipated good news for workers in the fields of medical practice/healthcare/hygiene in the world. Further, this invention renders a means for imparting a function and capability of enabling transmucosal administration and transdermal administration which are simpler than injection to biological drugs including conventional and future vaccines, toxoids, and the like and other various drugs.
  • the AD vehicles proposed by this invention are different from the conventional adjuvants used in the field of immunology in property and effect as described below.
  • the conventional adjuvants are ordinarily subcutaneously or intramuscularly inoculated to cause a local inflammation reaction and contain as an active ingredient a foreign matter that attracts antigen presentation cells and B- or T-lymphocytes to cause the cells to manifest their capabilities. Further, in order to maintain the inflammation reaction for a long time, a mineral oil and a metal salt that cause sustained release and retention of the antigen are used in combination.
  • Those known as the conventional mucosal vaccine/adjuvant are foreign matters such as the Escherichia coli bacteria heat-liable toxin and cholera toxin as described above and subject to a risk of causing harmful action and adverse reaction.
  • the AD vehicles according to this invention do not cause the local inflammation reaction.
  • the AD vehicles are derived from the biological component, and the active ingredients and active domains in the pulmonary surfactant are specified, thereby realizing effective mucosal vaccines by using the domains and low molecular peptides containing the domain regions. Therefore, the AD vehicles are remarkably safe and uninvasive.
  • Dry weights by wt% of Group A (pulmonary surfactant protein B and natural and synthetic peptides derived or originated from the protein B) and/or Group B (pulmonary surfactant protein C and natural and synthetic peptides derived or originated from the protein C) and Group C (lipid such as phospholipid and aliphatic acid) are as follows: about 0.1 to about 6.0 wt% of Group A; about 0.1 to about 6.0 wt% of Group B, and about 88 to about 99.8 wt% of Group C.
  • a mixer such as a box mixer [e.g. Vortex Mixer (trade name)].
  • lipid As the 96 mg of lipid (Group C), a mixture of 71 mg of phosphatidylcholine, 21 mg of phosphatidylglycerol, and 4 mg of phosphatidylserine or the like may be used (the total lipid amount is 96 mg). Also, in the case of using a pulmonary surfactant drug for RDS therapy which is reported to contain SP-B and SP-C, such as Surfacten and Surfacxin (artificial synthetic drug), it is possible to use a suspension prepared in accordance with the instruction attached thereto as it is as the AD vehicle solution.
  • a pulmonary surfactant drug for RDS therapy which is reported to contain SP-B and SP-C, such as Surfacten and Surfacxin (artificial synthetic drug
  • the antigen means a bacteria-derived antigen, a virus antigen, a toxoid, and the like that are highly purified to a purity of about 90% or more, an allergen such as those derived from cedar pollen or mite, a protein, a glycoprotein, a high molecular carbohydrate, nucleic acid, and the like.
  • an antigen mass an actual measurement value or a calculated value of a purity, a specific activity, a molecular amount, an antigen-antibody reaction of the antigen may be used.
  • a lipid amount or a phospholipid mass may be used in place of the AD vehicle amount since about 90 wt% of the AD vehicle is lipid or phospholipid in the typical preparation.
  • the dry weight (A) of the antigen in the mucosal vaccine of this invention may be about 0.1 to about 50 ⁇ g/kg, preferably about 0.3 to about 30 ⁇ g/kg.
  • a V/A for preferentially and selectively induce IgA production in such antigen amount may preferably be about 0.1 to about 1.0.
  • a V/A for inducing production of both of IgA and IgG may be in the range of about 1.0 to about 100, preferably about 20 to about 50.
  • a mucosal immunity vaccine obtained by coupling about 60% or more of the antigen to the AD vehicle in the above-described V/A ranges is capable of effectively inducing IgA antigen production and/or IgG antigen production.
  • lipids selected from 40 wt% or more of phospholipids such as phosphatidylcholine and dipalmitoylphosphatidylcholine, about 10 to 30 wt% of phosphatidylglycerol, about 2 to about 5 wt% of phosphatidylserine, about 1.0 to about 20 wt% of palmitic acid, and the like, a pulmonary surfactant-derived liposoluble (hydrophobic) protein, an active region of SP-B and/or SP-C or a synthetic peptide containing the active region, such as about 0.1 to about 12 wt% of a synthetic peptide formed from an active region of human SP-C (24-58 amino acid sequence set forth in SEQ ID NO: 20) FGIPCCPVHLKRLLIVVVVVVVLIVVVIVGALLMGL and prepared to achieve a total amount of the components of 100 w
  • a split influenza vaccine was prepared by the following operation. To the virus suspension dialyzed overnight with 0.004 M PBS [product of Takara Bio Inc. (Japan)], 0.05 vol% of ⁇ -propiolactone [product of Wako Pure Chemical Industries, Ltd. (Japan)] was added to achieve a final concentration of 8 nM, followed by virus inactivation by incubation in an ice bath for 18 hours.
  • ⁇ -propiolactone was subjected to hydrolytic degradation by incubation at 37°C for 1.5 hours.
  • Tween 80 [product of Wako Pure Chemical Industries, Ltd. (Japan)] was added to achieve a final concentration of 0.1 wt%
  • diethylether product of Wako Pure Chemical Industries, Ltd. (Japan)] in an amount equivalent to that of Tween 80 was added, followed by inverted mixing at 4°C for 2 hours.
  • the thus-obtained liquid was centrifuged at 2,000 rpm for 5 minutes, followed by collection of a supernatant thereof.
  • the split influenza vaccine stock solution obtained by Experimental Example 1 and the AD vehicle described in Reference Example 1 were mixed. After suspending the AD vehicle into PBS (phosphate buffer saline) until a concentration required for vaccination is achieved, a supersonic wave treatment was performed at a room temperature for 5 minutes to obtain a uniform suspension. To the suspension, 0.1 ⁇ g (dry weight) of the vaccine stock solution was added with respect to 0.1 ⁇ g (dry weight) of the AD vehicle, followed by mixing with Vortex Mixer, and the mixture was left to stand at a room temperature for one hour to prepare a transnasal/mucosal vaccine for IgA induction.
  • PBS phosphate buffer saline
  • transnasal/mucosal vaccine for both of IgA and IgG induction was prepared in the same manner as described above.
  • the vaccine obtained by Experimental Example 2 was diluted with PBS to achieve a vaccine concentration of 0.1 ⁇ g/ ⁇ l (dry weight), and 1 ⁇ l of the diluted vaccine was dropped into each of nostrils of each of the etherized mice to administer 2 ⁇ l of the diluted vaccine to each of the mice.
  • the same amount (2 ⁇ l) of PBS was administered by nasal drip.
  • Secondary immunization was performed by the nasal drip at 4 weeks after the primary immunization, and a nasal wash, an alveolar wash, a blood serum of each of the mice were prepared at 2 weeks after the secondary immunization to be used as specimens for measurements of both of IgA and IgG antibodies specific to the vaccine strain virus.
  • mice Each of the vaccinated mice was subjected to thoracotomy and laparotomy under pentobarbital anesthesia, and the trachea was cut to insert an atom knotted vein catheter 3Fr [product of Atom Medical Corporation (Japan)] into the lung, followed by injection of 1 ml of a normal saline solution and collection of the wash. This operation was repeated for three times to obtain 3 ml of washes to be used as an alveolar wash. After collecting the lung wash, an atom vein catheter was inserted from the opened trachea toward the nostrils, and 1 ml of a normal saline solution was injected, and a liquid flowed out of the nostrils was collected. This liquid was used as a nasal wash. Further, blood from the heart was centrifuged at 5,000 rpm for 10 minutes to obtain a supernatant, and the supernatant was used as a blood serum.
  • 3Fr product of Atom Medical Corporation (Japan)
  • a protein content of each of the nasal wash, the lung wash, and the blood serum was measured by using BCA Protein Assay Reagent Kit [product of Peirce (USA)] ( Anal. Biochem., 150, 76-85, 1985 ). Absorbance at 562 nm was measured by using SPECTRA Max PLUS 384 (product of Molecular Devices Corporation (USA)).
  • the IgA and IgG antibodies were purified and prepared in the manner described below.
  • affinity chromatography using recombinant Escherichia coli bacteria-expressed protein G sepharose 4B column [product of Zymed Laboratories Inc. (USA)] an IgG fraction was purified from the alveolar wash of each of the influenza-vaccinated and influenza virus-infected mice.
  • an anti-mouse IgA goat IgG antibody [product of Sigma (USA)] was coupled to a BrCN activated sepharose 4B column [product of Amersham Bioscience (USA)], and an IgA fraction was purified from a flow-through fraction of Protein G by means of affinity chromatography using the column.
  • an antigen affinity chromatography in which the inactivated split influenza vaccine antigen used for the immunization was coupled to a BrCN activated sepharose column, was employed for purification/preparation of each of IgA and IgG antibodies specific to the influenza virus.
  • a coupling reaction was performed by using a 0.1 M NaHCO 3 /0.5 M NaCl buffer solution (pH 8.5), and the free ligand was eliminated by using 1 M acetic acid/0.5 M NaCl (pH 8.5) and neutralized with PBS (pH 7.5).
  • Each of the affinity chromatographies was performed in such a manner that the affinity coupling reaction and free antibody elimination conducted by using PBS (pH 7.5) followed by elution of the specific antibodies by using a glycine-HCI buffer solution (pH 2.8).
  • the eluted fraction was immediately neutralized with 0.5 M Tris-HCl buffer solution (pH 9.0) and dialyzed with Milli Q water, followed by freeze-drying.
  • the IgA and IgG were dissolved into PBS in each use as standard reagents (antibodies) for ELISA.
  • a content of each of the anti-influenza virus IgA and IgG antibodies in the nasal wash, the alveolar wash, and the blood serum was determined by ELISA.
  • ELISA was performed by using Mouse ELISA quantitatin kit [product of Bethyl Laboratories (USA)]. After adding 1 ⁇ g of the vaccine and 100 ⁇ g of a PBS solution of a 1 ⁇ g/ml-cow serum albumin [BSA; product of Sigma (USA)] to each of wells of a 96 well Nunc imunoplate [product of Nalgen Nunc International (USA)], an immobilization reaction was performed at 4°C overnight.
  • BSA 1 ⁇ g/ml-cow serum albumin
  • rinsing with a cleaning liquid 50 mM Tris, 0.14 M NaCl, 0.05 wt% Tween 20, pH 8.0
  • a cleaning liquid 50 mM Tris, 0.14 M NaCl, 0.05 wt% Tween 20, pH 8.0
  • 200 ⁇ l of 50 mM Tris-HCL buffer solution (pH 8.0) containing 0.15 M NaCl and 1 wt% BSA was added to each of the wells to allow a blocking reaction at a room temperature for one hour.
  • sample coupling buffer solution 50 mM Tris, 0.15 M NaCl, 0.05 wt% Tween 20, 1 wt% BSA, pH 8.0
  • An antibody mouse IgA goat IgG antibody or IgG-horse radish peroxidase [product of Bethyl Laboratories Inc.
  • Surfacten [product of Mitsubishi Pharma Corporation (Japan)] was used as the AD vehicle, and a mixture of Surfacten and the influenza inactivated vaccine obtained by Example 1 was subjected to supersonic wave treatment to prepare a mucosal vaccine. Surfacten was suspended in use into PBS to achieve a concentration required for vaccination, and a uniform suspension was obtained by a supersonic wave treatment at a room temperature for one minute.
  • the Surfacten suspension was added to 0.2 ⁇ g (dry weight) of the vaccine in such a manner that concentrations of the Surfacten by way of a phospholipid amount thereof became 0 (no addition), 0.02 ⁇ g, 0.1 ⁇ g, 0.2 ⁇ g, 1.0 ⁇ g, and 2.0 ⁇ g, followed by mixing by Vortex Mixer and supersonic wave treatment at a room temperature for 3 minutes.
  • the vaccine liquids were incubated at a room temperature for one hour before administration.
  • each of the vaccine liquids was diluted with PBS to obtain a solution containing the vaccine in an amount equivalent to 0.1 ⁇ g/ ⁇ l by dry weight, and the diluted vaccine liquid was administered to nostrils of BALB/c mouse for primary immunization.
  • a secondary immunization was performed in the same manner as in the primary immunization at 4 weeks after the primary immunization, and a nasal wash, an alveolar wash, and a blood serum were collected and prepared from each of the mice at 2 weeks after the secondary immunization to be used for quantitative detection of anti-influenza IgA and IgG.
  • the amount of Surfacten added to the vaccine was 0.2 ⁇ g or less (hereinafter referred to as low dose group)
  • the anti-influenza IgA production amounts at the nasal mucosa and alveolar mucosa were increased depending on the Surfacten amount ( Figs. 1 and 2 ).
  • enhancement associated with the increase in Surfacten was not observed with the IgG production amount in the low dose group, and productions of a blood antibody, IgA, and IgG were not induced ( Fig. 3 ).
  • the Surfacten amount was 1.0 to 2.0 ⁇ g (hereinafter referred to as high dose group)
  • the anti-influenza IgG production amounts at the nasal mucosa and the alveolar mucosa were increased depending on the Surfacten amount (see Figs. 1 and 2 ).
  • the blood IgG production amount was significantly increased in Group in which Surfacten was used ( Fig. 3 ).
  • a significant increase in IgA production amount was not observed as compared to that of the low dose group, and enhancement of mucosal immunity effect of the low dose group was not observed ( Figs. 1 and 2 ).
  • V/A weight ratio of the phospholipid amount/dry vaccine amount in the AD vehicle
  • the phospholipid mass (weight) in the AD vehicle was used for the value V in the V/A calculation.
  • the lipid amount (including phospholipid) or the AD vehicle amount (dry weight) was used as the value V in adjustment of the weight ratio V/A when carrying out this invention.
  • the synthetic peptide FGIPCCPVHLKRLLIVVVVVVLIVVVIVGALLMGL [synthesized by Greiner Bio-One (Germany)] of an active region of human SP-C having purity of 95% or more was ordered/purchased.
  • a synthetic SP-C suspension was obtained by suspending the synthetic peptide into a CM suspension (buffer solution obtained by mixing chloroform and ethanol at a volumetric ratio of 2: 1) to achieve a protein amount thereof of 10 mg/ml, and the synthetic SP-C suspension was used for preparation of an artificial AD vehicle (artificial human pulmonary surfactant) described below.
  • Trifluoroacetic acid (TFA) in an amount same as that of the SP-C suspension was added to the SP-C suspension to be perfectly dissolved into the SP-C suspension.
  • TFA Trifluoroacetic acid
  • the mixture was dried by using a rotary evaporator at 40°C.
  • the dried mixture was suspended into a 10%-ethanol solution until the phospholipid amount becomes 10 mg/ml, and the suspension was mixed by shaking in a hot water bath of 45°C for 15 minutes. After freeze-drying the mixture, the mixture was stored as an artificial dried human pulmonary surfactant (synthetic AD vehicle) at -30°C to -4°C.
  • the above-described artificial dried AD vehicle was suspended into PBS to achieve a phospholipid amount of 30 mg/ml, followed by a supersonic wave treatment at a room temperature for one minute to obtain a uniform suspension.
  • the artificial surfactant or Surfacten (described in Example 1) as a positive control of immunization induction effect was added to the suspension to achieve 2.0 ⁇ g of the phospholipid amount with respect to 0.2 ⁇ g of the vaccine obtained by Example 1, followed by mixing by Vortex Mixer and a supersonic wave treatment at a room temperature for 3 minutes.
  • the thus-obtained transnasal mucosal vaccine was incubated for one hour before administration.
  • the vaccine liquid was diluted with PBS to obtain a 0.1 ⁇ g/ ⁇ l solution of the vaccine to be transnasally administered.
  • each of BALB/c mice was immunized, and a nasal wash, an alveolar wash, and a blood serum were collected/prepared from each of the mice at 2 weeks after the secondary immunization to be used for quantitative determination of anti-influenza IgA and IgG.
  • levels of anti-influenza IgA and IgG were measured by quantitative determination by ELISA, and influences and effects to be exerted on induction of mucosal immunity and/or humoral immunity by the artificial human pulmonary surfactant (synthetic AD vehicle) containing the synthetic SP-C as the AD vehicle in the transnasal/mucosal vaccine were evaluated based on results of significant difference test of the levels.
  • synthetic AD vehicle synthetic SP-C as the AD vehicle in the transnasal/mucosal vaccine
  • an IgA production amount of the artificial surfactant-mixed vaccine administration group was significantly increased to a level almost identical with that of the positive control Surfacten-mixed vaccine administration group. Also, IgG production that had not been detected in the AD vehicle-free vaccine group was detected ( Fig. 4 ).
  • an IgG production amount of the artificial surfactant-mixed vaccine administration group was significantly increased like that of the positive control Surfacten-mixed vaccine administration group. Also, though IgA production was detected, no significant difference between the production amount and that of the AD vehicle-free vaccine administration group was observed. However, since no significant difference was observed between the IgA production and the Surfacten-mixed vaccine administration group, it was evaluated that the IgA production induction capacity of the artificial surfactant-mixed vaccine was similar to that of the positive control Surfacten-mixed vaccine ( Fig. 5 ).
  • an IgG production amount of the artificial surfactant-mixed vaccine administration group was significantly increased to a level almost identical with that of the positive control Surfacten-mixed vaccine administration group ( Fig. 6 ).
  • the artificial synthetic surfactant has induction capability for both of mucosal immunity and humoral immunity like the cow-derived Surfacten.
  • Surfacten (ST) was purchased from Mitsubishi Pharma Corporation.
  • Influenza Split Vaccine A/New York/55/2004 (H3N2), A/New Caledonia/20/99 (H1N1), B/Shanghai/361/2002 were obtained from Osaka University Research Institute for Microbial Disease.
  • a synthetic surfactant Preparation substrate DPPC (dipalmitoylphosphatidylcholine) was obtained from Wako Pure Chemical Industries, Inc. and Sigma; E-PG (egg-phosphatidylglycerol) was obtained from Avanti and Sigma; and PA (palmitic acid) was obtained from Wako Pure Chemical Industries, Inc.
  • DPPC dipalmitoylphosphatidylcholine
  • E-PG egg-phosphatidylglycerol
  • PA palmitic acid
  • Synthetic peptides were obtained from Gliner, Hayashi Kasei Co., Ltd., and the like.
  • the synthetic peptides were SP-B(1-25) (SEQ ID NO: 21), SP-B(20-60) (SEQ ID NO: 22), SP-B(64-80) (SEQ ID NO: 23), SP-B(1-60) (SEQ ID NO: 24), SP-C(1-19) (SEQ ID NO: 25), synthetic peptide KL-4 which is under development and study as a respiratory disorder therapeutic drug (SEQ ID NO: 26), SP-CL(7-28) (SEQ ID NO: 27), SP-C33 (SEQ ID NO: 28), SP-C(FFI) (SEQ ID NO: 29), and SP-C(KLS) (SEQ ID NO: 30).
  • BCATM Protein Assay Kit was purchased from PIERCE, and Phospholipid C Test Wako was purchased from Wako Pure Chemical Industries, Inc.
  • Each of the synthetic peptides was dissolved into an appropriate solvent.
  • the hardly-soluble synthetic peptide was dissolved into TFA at a concentration of 5 to 20 mg/ml.
  • DPPC, e-PG, and a PA lipid mixture (75:25:10; w/w/w) (hereinafter referred to as three-lipid mixture liquid) dissolved into a chloroform/methanol (2:1, v/v) mixture liquid was added to the synthetic peptide (0.6 to 2.0 %(mol) with respect to phospholipid (PL)) dissolved into TFA.
  • the mixture liquid was dried under a reduced pressure by a decompression concentrator.
  • a 10%-ethanol solution was added to the dried matter, and pH thereof was adjusted to 6 to 7 with a N-NaOH solution, followed by heating at 42°C to 45°C for 3 to 10 minutes. After cooling, the concentration of the mixture was adjusted to 1 to 10 mg P1/mL, and 1 to 5 mg PL equivalent amount was distributed and freeze-dried.
  • the thus-obtained freeze-dried matter i.e. the synthetic AD vehicle, was suspended into a normal saline solution to be used for a test for coupling to the vaccines.
  • AD vehicles synthetic surfactant: synthetic AD vehicle ST
  • SSF-28, SSF-30, SSF-24, AND SSF-43 were prepared by using the synthetic peptide SP-B(1-60), SP-B(20-60), SP-B(64-80), and SP-C(1-19), respectively.
  • SSF-45, SSF-44, SSF-46, SSF-47, and SSF-48 were prepared by using KL-4, SP-CL(7-27), SP-C33, SP-C(FFI), and SP-C(KLS), respectively.
  • the synthetic peptides SP-B(64-80) and SP-C(1-35) were dissolved into TFA at a concentration of 5 mg/mL.
  • the three-lipid mixture was added to the synthetic peptides dissolved into TFA in an amount of 0.6% (mol) with respect to phospholipid of the three-lipid mixture. After that, the operation described above was performed to prepare SSF-41.
  • a suspension test was performed by setting: an AD vehicle concentration to 0.5 mg/mL, a vaccine concentration to 0.05 mg/mL, a ratio between the concentrations to 10:1, and a liquid amount to 0.4 mL scale.
  • a normal saline solution was added to ST to achieve a concentration of 30 mg/mL, followed by pipetting to obtain a suspension.
  • 7 ⁇ L of the suspension was withdrawn into an Eppendorf tube, and 382 ⁇ L of a normal saline solution was added thereto, followed by a supersonic wave treatment by using Digital SONIFIER 250D (product of Branson) at an ice cool temperature for one minute.
  • 11 ⁇ L of a vaccine solution (A/New Caledonia/20/99(H1N1), 1.78 mg Prot/mL) was added to the suspension, followed by a supersonic wave treatment at an ice cool temperature for 3 minutes. After that, the solution was left to stand at a room temperature for 2 hours, during which inverted mild mixing was performed every 30 minutes for 3 to 5 times.
  • the AD vehicle ST-Vaccine mixture was centrifuged by using High Speed Refrigated micro centrifuge (Tomy MX 301) at a temperature of 4°C to 5°C at 20,000 ⁇ g for 15 minutes. From the thus-obtained supernatant, 25 ⁇ L was separated to be used for measurement of a vaccine amount as a protein mass by using BCATM Protein Assay Kit.
  • Ratios of coupling of the three vaccines (A/New York, A/new Caledonia, and B/Shanghai) to the AD vehicle ST were 60% to 70%.
  • AD vehicle ST of (1) synthetic AD vehicles SSF-14, SSF-30, and SSF-24 were used. Also, the vaccine used for the test was A/New Caledonia.
  • a coupling test was performed by setting: a synthetic AD vehicle concentration (calculated in terms of phospholipid measured using Phosphlipid C Test Wako (Wako Pure Chemical Industries, Ltd.; 433-36201) to 2.5 mg/mL, a vaccine concentration to 0.05 mg/mL, a ratio between the concentrations to 50:1, and a liquid amount to 0.4 mL scale and in accordance with (1).
  • Results are shown in Table 1. A coupling ratio of the vaccine to SSF-14 was just under 60%, and coupling ratios of SSF-24 and SSF-30 were 90% or more.
  • a coupling test was performed by setting: a synthetic AD vehicle concentration (calculated in terms of phospholipid) to 0.5 mg/mL, a vaccine concentration to 0.05 mg/mL, a ratio between the concentrations to 10:1, and a liquid amount to 0.4 mL scale and in accordance with (1).
  • Results are shown in Table 2. A coupling ratio of the vaccine to SSF-43 was about 80%.
  • a coupling test was performed by using SSF-28 as the synthetic AD vehicle and using A/New Caledonia/20/99(H1N1), A/New York/55/2004(H3N2), and B/Shanghai/361/2002 as the vaccines.
  • the nasal mucosa lymphatic tissue varies greatly depending on animal specie. Though a multiple of lymphatic tissues corresponding to Peyer's Patches of the intestine exists at the nasal mucosa of mice and rats, such lymphatic tissue does not exist at the human nasal mucosa, and it is difficult to mention that the experimental results of the transnasal mucosal vaccine for mice and rats are directly applicable to human.
  • the lymphatic tissue of the human nostrils and pharynx consists mainly of Waldeyer's ring formed of the lymphatic tissues of tonsils and adenoid, and the animal having the lymphatic tissue relatively similar to the tissue is a pig. Accordingly, based on the results of the AD vehicle obtained on mice, the AD vehicle effect was investigated by using minipigs as a preclinical test assuming a human therapy to detect an administration amount, an administration method, and an adverse effect of the AD vehicle in the clinical test.
  • Postweaning Minipigs of 1 to 2 month-year-old (weight: 2.2 to 6.6 kg) were used. Three subjects were used for one group, with which absence of abnormality was confirmed by testing 22 kinds of viruses, bacteria, parasites, and the like with which pigs are ordinarily infected.
  • a split antigen of A/New Caledonia/20/99(H1N1) was used.
  • Surfacten TM (AD vehicle ST) purchased from Mitsubishi Pharma Corporation was used.
  • BCMTM Protein Assay Kit was purchased from PIERCE, and Phospholipid C Test Wako was purchased from Wako Pure Chemical Industries, Inc.
  • Vaccine inoculation, blood collection, and nasal mucus collection were conducted under anesthesia.
  • weights of the minipigs were measured in advance of the anesthesia, and Domitol (product of Meiji Seika Kaisha, Ltd.) and Dormicam (product of Astellas Pharma Inc.) were mixed to be used for the anesthesia.
  • Blood was collected from the venous cavity. Collection of nasal mucus was conducted by wiping the interior of the right and left nostrils with a sterilized cotton swab and dipping the cotton swab into 2 ml of a normal saline solution, followed by squeezing the cotton swab. A part of the nasal mucus was immediately subjected to cytopathologic diagnosis to investigate presence/absence of an inflammation reaction in the nostrils.
  • Weights and body temperatures were measured once a week, and health states were recorded twice a day by visual inspection.
  • a measurement of an antigen specific antibody was conducted in accordance with the antigen specific antibody measurement in mouse and by employing an enzyme antibody technique (ELISA) using anti-pig IgA and IgG secondary antibodies.
  • ELISA enzyme antibody technique
  • the split antigen of influenza A/New Caledonia/20/99(H1N1) was coupled to the AD vehicle ST to be suspended into 100 ⁇ l of a normal saline solution.
  • the suspension was administered in the form of nasal drops to the minipigs (NIBS) with which no abnormality was detected by an ordinary pathogen test to measure production amounts of the antibodies (IgA, IgG) specific to the vaccine.
  • a booster administration was performed in such a manner that an administration in the same manner as the initial administration was performed at 4 weeks after the initial administration, and the measurement was ended at 2 weeks after the booster administration.
  • Antigen amounts were detected in 5 groups of 0.3 ⁇ g/Kg, 1.5 ⁇ g/Kg, 4.5 ⁇ g/Kg, 15 ⁇ g/Kg, and 30 ⁇ g/Kg.
  • a nasal mucus was measured for evaluating a local mucosal immunity, and a blood antibody production amount was measured for evaluating a systemic immunity.
  • the nasal mucus and the blood IgA and IgG values were scarcely changed after the 0.3 to 30 ⁇ g/Kg transnasal administration of the split influenza antigen alone or slightly increased to be short of the effective infection prevention antibody concentration (equal to or more than 40 antibodies of antigen-specific IgG/IgA).
  • the transnasal administration of the suspension wherein the split influenza antigen and the AD vehicle ST are coupled to each other by adding to the antigen the AD vehicle ST as an adjuvant in an amount (weight ratio) 10 times of the antigen the IgA-specific antibody was particularly induced in the nasal mucus, while the IgG-specific antibody was particularly induced in the blood, depending on the administered antigen amount (0.3 to 30 mg/Kg).
  • the antigen-specific antibodies were induced to an antibody value of 200 to 800 depending on the administered antigen amount, and, thus, the effective adjuvant effect of the AD vehicle ST was proved.
  • the IgA antibody value and IgG antibody value of the vaccine-specific antibodies defined herein was obtained by measuring the dilution degree that caused the value before the vaccine administration by setting an average of measurement values indicated as a result of the reaction between the vaccine antigen and the nasal mucus/blood before the vaccine administration as a reference, and the thus-detected dilution degree was considered as the antigen-specific IgA IgG antibody values.
  • the IgA was preferentially produced, and the IgA antibody value was 5 to 8 times of the IgG antibody value. Increase tendencies of the IgA antibody value and the IgG antibody value depending on the antigen amount were not different from each other by a large scale.
  • the production of the antibodies in the nasal mucus after the initial administration was weak and considerably increased by the booster administration.
  • IgG in the nasal mucus with the single use of the split influenza antigen was increased after the booster administration to a value that was about ten times that before the booster administration though the increase was relatively weak.
  • the IgG antibody value was 1 to 1.6 times of the IgA antibody value to reveal that IgG is preferentially produced in the blood. Both of the antibodies were increased to the antibody values of 100 to 200 after the primary immunization, and both of the IgA and IgG antibody values after the booster administration were 4 times of the values before the booster administration. Blood IgG with the single use of the split influenza antigen was increased after the booster administration to a value that was about ten times that before the booster administration though the increase was relatively weak.
  • the AD vehicle according to this invention enables safe administration or inoculation of an arbitrary vaccine antigen, a toxoid, an allergen, a drug, and the like which is performed transmucosally, transdermally, or the like.
  • a weight ratio V/A between the AD vehicle amount (V) and a substance to be mounted on the AD vehicle, such as the antigen amount (A) it is possible to adjust an antibody production induction function of the AD vehicle. More specifically, the AD vehicle preferentially and selectively induces IgA production when the V/A is about 1 or less and induces production of both of IgA and IgG when the V/A is adjusted to exceed 1.
  • the AD vehicle is widely useful as a vehicle or as a means for delivery and transportation of an active ingredient of a transnasal/mucosal vaccine, a therapeutic/prophylactic agent for allergy, and the like and is usable in the industrial fields of pharmaceutical agents according to vaccine and allergy therapy, veterinary, fishery, and the like.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Immunology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Virology (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pulmonology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Medicinal Preparation (AREA)
EP06782373.2A 2005-08-05 2006-08-04 VEHICULE POUR ANTIGENES ET MEDICAMENTS PERMETTANT DE BASCULER DE LA PRODUCTION SELECTIVE D'ANTICORPS IgA A LA PRODUCTION D' ANTICORPS TANT IgA QUE IgG, ET VACCIN TRANSNASAL/TRANSMUCOSIQUE UTILISANT LE VEHICULE Ceased EP1930025B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2005227504 2005-08-05
PCT/JP2006/315515 WO2007018152A1 (fr) 2005-08-05 2006-08-04 VÉHICULE POUR ANTIGÈNES ET MÉDICAMENTS PERMETTANT DE BASCULER DE LA PRODUCTION SÉLECTIVE D'ANTICORPS IgA À LA PRODUCTION D’ANTICORPS TANT IgA QUE IgG, ET VACCIN TRANSNASAL/TRANSMUCOSIQUE UTILISANT LE VÉHICULE

Publications (3)

Publication Number Publication Date
EP1930025A1 true EP1930025A1 (fr) 2008-06-11
EP1930025A4 EP1930025A4 (fr) 2008-12-03
EP1930025B1 EP1930025B1 (fr) 2013-05-01

Family

ID=37727335

Family Applications (1)

Application Number Title Priority Date Filing Date
EP06782373.2A Ceased EP1930025B1 (fr) 2005-08-05 2006-08-04 VEHICULE POUR ANTIGENES ET MEDICAMENTS PERMETTANT DE BASCULER DE LA PRODUCTION SELECTIVE D'ANTICORPS IgA A LA PRODUCTION D' ANTICORPS TANT IgA QUE IgG, ET VACCIN TRANSNASAL/TRANSMUCOSIQUE UTILISANT LE VEHICULE

Country Status (5)

Country Link
US (1) US8211442B2 (fr)
EP (1) EP1930025B1 (fr)
JP (1) JP5028627B2 (fr)
CN (1) CN101257919B (fr)
WO (1) WO2007018152A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8287887B2 (en) 2008-04-02 2012-10-16 The University Of Tokushima Antigen-and-drug vehicle comprising synthetic peptide, and mucosal vaccine using the same
US20130045232A1 (en) * 2010-03-02 2013-02-21 Hiroshi Kido Mucosal vaccines
EP3201341A4 (fr) * 2014-10-01 2018-06-27 Plant Health Care, Inc. Peptides eliciteurs de reponse hypersensible et leur utilisation
US10524472B2 (en) 2014-10-01 2020-01-07 Plant Health Care, Inc. Elicitor peptides having disrupted hypersensitive response box and use thereof
US10793608B2 (en) 2016-04-06 2020-10-06 Plant Health Care, Inc. Hypersensitive response elicitor-derived peptides and use thereof
US11371011B2 (en) 2016-04-06 2022-06-28 Plant Health Care, Inc. Beneficial microbes for delivery of effector peptides or proteins and use thereof

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110014230A1 (en) 2008-03-18 2011-01-20 Novartis Ag preparation of influenza virus vaccine antigens
US8785390B2 (en) 2009-01-30 2014-07-22 Alphabeta Ab Methods for treatment of Alzheimer's disease
JP5893614B2 (ja) 2010-06-24 2016-03-23 アルファベータ・エービーＡｌｐｈａＢｅｔａ ＡＢ アルツハイマー病および家族性認知症の治療のための化合物および方法
CN101948526B (zh) * 2010-09-02 2012-12-05 华东理工大学 一种肺泡表面活性蛋白质(sp-b和sp-c)的分离纯化方法
US20130243817A1 (en) * 2010-09-30 2013-09-19 Eurocine Vaccines Ab Vaccine compositions
US9522170B2 (en) 2011-04-05 2016-12-20 Alphabeta Ab Methods of screening compounds for the fibril formation of Aβ peptides based on a decreased trimer/monomer ratio of a chaperone protein
EP2752199A4 (fr) 2011-08-29 2015-04-29 Univ Tokushima Vaccin muqueux contre rsv
CN104379170B (zh) 2012-06-20 2017-05-03 国立大学法人东京大学 粘膜免疫刺激剂及hpv感染症治疗用经口药物组合物
EP2742952A1 (fr) 2012-12-17 2014-06-18 Eurocine Vaccines AB Composition vaccinale
EP3820510A4 (fr) * 2018-07-10 2022-05-04 Seqirus Pty Ltd Élimination d'agglomérats
US20210338809A1 (en) 2018-08-30 2021-11-04 Tokushima University Immune Tolerance-Inducing Agent and Therapeutic or Prophylactic Agent for Allergic Disorder

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0652011A1 (fr) * 1992-06-24 1995-05-10 Tokyo Tanabe Company Limited Agent prophylactique et agent curatif contre des maladies virales des voies respiratoires
WO2000006198A2 (fr) * 1998-07-30 2000-02-10 Id-Lelystad Adjuvant comprenant un tensioactif pulmonaire
US20020155151A1 (en) * 1997-02-11 2002-10-24 Franz Enzmann Transdermal, oral and intravenous preparations of 2,3-dimethoxy-5-methyl-6-decaprenyl-1,4-benzoquinone
WO2005097182A1 (fr) * 2004-04-05 2005-10-20 The University Of Tokushima Vehicule de medicament antigenique permettant l'administration transmuqueuse et transdermqiue et procédé d'induction d'immunité muquese, vaccins par voie muqueuse et srm

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03135923A (ja) 1989-10-20 1991-06-10 Chemo Sero Therapeut Res Inst 百日咳毒素ｂオリゴマーの構成サブユニットを含有する経鼻接種ワクチン
AUPM399594A0 (en) 1994-02-21 1994-03-17 Csl Limited Antigenic preparation for treatment or prevention of helicobacter infection
EP0919243A1 (fr) 1997-11-25 1999-06-02 Duphar International Research B.V Vaccin dont l'adjuvant est constitué de sous-unités B de l'entérotoxine thermolabile (LTB) d'Escherichia coli
US6248329B1 (en) * 1998-06-01 2001-06-19 Ramaswamy Chandrashekar Parasitic helminth cuticlin nucleic acid molecules and uses thereof
JP2002526425A (ja) 1998-10-05 2002-08-20 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 粘膜免疫を刺激するための方法およびアジュバント
AUPQ797700A0 (en) * 2000-06-06 2000-06-29 Austin Research Institute, The Vaccine
JP2002050452A (ja) 2000-08-02 2002-02-15 Canon Inc 加熱体、像加熱装置および画像形成装置
JP2003050452A (ja) 2001-08-06 2003-02-21 Fuji Photo Film Co Ltd 定着用添加剤組成物および、該添加剤組成物と定着剤濃縮組成物とを用いたハロゲン化銀写真感光材料の処理方法
JP2003116385A (ja) 2001-10-15 2003-04-22 Univ Of The Ryukyus 日本脳炎ワクチンをコードする遺伝子を含むトランスジェニック植物
WO2003035679A2 (fr) * 2001-10-25 2003-05-01 Medical Research Council Molecules

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0652011A1 (fr) * 1992-06-24 1995-05-10 Tokyo Tanabe Company Limited Agent prophylactique et agent curatif contre des maladies virales des voies respiratoires
US20020155151A1 (en) * 1997-02-11 2002-10-24 Franz Enzmann Transdermal, oral and intravenous preparations of 2,3-dimethoxy-5-methyl-6-decaprenyl-1,4-benzoquinone
WO2000006198A2 (fr) * 1998-07-30 2000-02-10 Id-Lelystad Adjuvant comprenant un tensioactif pulmonaire
WO2005097182A1 (fr) * 2004-04-05 2005-10-20 The University Of Tokushima Vehicule de medicament antigenique permettant l'administration transmuqueuse et transdermqiue et procédé d'induction d'immunité muquese, vaccins par voie muqueuse et srm

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO2007018152A1 *

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8287887B2 (en) 2008-04-02 2012-10-16 The University Of Tokushima Antigen-and-drug vehicle comprising synthetic peptide, and mucosal vaccine using the same
US20130045232A1 (en) * 2010-03-02 2013-02-21 Hiroshi Kido Mucosal vaccines
US9540420B2 (en) * 2010-03-02 2017-01-10 Tokushima University Mucosal vaccines
US10918104B2 (en) 2014-10-01 2021-02-16 Plant Health Care, Inc. Elicitor peptides having disrupted hypersensitive response box and use thereof
US10856547B2 (en) 2014-10-01 2020-12-08 Plant Health Care, Inc. Hypersensitive response elicitor peptides and use thereof
US10524472B2 (en) 2014-10-01 2020-01-07 Plant Health Care, Inc. Elicitor peptides having disrupted hypersensitive response box and use thereof
US10524473B2 (en) 2014-10-01 2020-01-07 Plant Health Care, Inc. Elicitor peptides having disrupted hypersensitive response box and use thereof
US10743538B2 (en) 2014-10-01 2020-08-18 Plant Health Care, Inc. Elicitor peptides having disrupted hypersensitive response box and use thereof
US11820992B2 (en) 2014-10-01 2023-11-21 Plant Health Care, Inc. Hypersensitive response elicitor peptides and use thereof
US10856546B2 (en) 2014-10-01 2020-12-08 Plant Health Care, Inc. Hypersensitive response elicitor peptides and use thereof
US10470461B2 (en) 2014-10-01 2019-11-12 Plant Health Care, Inc. Hypersensitive response elicitor peptides and use thereof
US10897900B2 (en) 2014-10-01 2021-01-26 Plant Health Care, Inc. Hypersensitive response elicitor peptides and use thereof
EP3201341A4 (fr) * 2014-10-01 2018-06-27 Plant Health Care, Inc. Peptides eliciteurs de reponse hypersensible et leur utilisation
US11820797B2 (en) 2014-10-01 2023-11-21 Plant Health Care, Inc. Elicitor peptides having disrupted hypersensitive response box and use thereof
US11725027B2 (en) 2016-04-06 2023-08-15 Plant Health Care, Inc. Hypersensitive response elicitor-derived peptides and use thereof
US11371011B2 (en) 2016-04-06 2022-06-28 Plant Health Care, Inc. Beneficial microbes for delivery of effector peptides or proteins and use thereof
US10793608B2 (en) 2016-04-06 2020-10-06 Plant Health Care, Inc. Hypersensitive response elicitor-derived peptides and use thereof

Also Published As

Publication number Publication date
JP5028627B2 (ja) 2012-09-19
EP1930025B1 (fr) 2013-05-01
US20090130131A1 (en) 2009-05-21
EP1930025A4 (fr) 2008-12-03
JPWO2007018152A1 (ja) 2009-02-19
WO2007018152A1 (fr) 2007-02-15
CN101257919A (zh) 2008-09-03
CN101257919B (zh) 2012-08-29
US8211442B2 (en) 2012-07-03

Similar Documents

Publication Publication Date Title
EP1930025B1 (fr) VEHICULE POUR ANTIGENES ET MEDICAMENTS PERMETTANT DE BASCULER DE LA PRODUCTION SELECTIVE D'ANTICORPS IgA A LA PRODUCTION D' ANTICORPS TANT IgA QUE IgG, ET VACCIN TRANSNASAL/TRANSMUCOSIQUE UTILISANT LE VEHICULE
JP4817625B2 (ja) 粘膜免疫誘導アジュバントを含む新規ワクチン
AU2006304895B2 (en) Canine influenza virus and related compositions and methods of use
JP5730862B2 (ja) オクロバクトラム・インターメディウムのリポ多糖類および哺乳動物の免疫刺激剤としてのそれらの使用
KR20140043348A (ko) 리포솜 제제
JP2009523722A (ja) ポリイノシン酸−ポリシチジル酸を基礎としたアジュバントを含む粘膜免疫原性物質
EP2543387B1 (fr) Vaccin muqueux
JP4843792B2 (ja) 経粘膜及び経皮投与を可能にする抗原薬物ヴィークル、これを用いる粘膜免疫の誘導方法、粘膜ワクチン及びｄｄｓ
EP2281574B1 (fr) Véhicule antigène-et-médicament comprenant un peptide synthétique, et vaccin par voie muqueuse utilisant celui-ci
JP2013506682A (ja) 異種亜型インフルエンザｔ細胞応答を誘発するためのペプチド
JPWO2013085021A1 (ja) 抗体産生増強用の組成物
CA3076263C (fr) Hemagglutinine synthetique en tant que vaccin universel contre une infection par des virus de la grippe de type b (ibv)
JP2001151698A (ja) インフルエンザワクチン
WO2021206103A1 (fr) Vaccin pour une administration sous-cutanée
US9463236B2 (en) RSV mucosal vaccine
MX2008008967A (en) Mucosal immunogenic substances comprising a polyinosinic acid - polycytidilic acid based adjuvant

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20080305

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): DE FR GB IT

RBV Designated contracting states (corrected)

Designated state(s): DE FR GB IT

A4 Supplementary search report drawn up and despatched

Effective date: 20081105

17Q First examination report despatched

Effective date: 20090414

DAX Request for extension of the european patent (deleted)
REG Reference to a national code

Ref country code: DE

Ref legal event code: R079

Ref document number: 602006036063

Country of ref document: DE

Free format text: PREVIOUS MAIN CLASS: A61K0039000000

Ipc: A61K0038170000

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

RIC1 Information provided on ipc code assigned before grant

Ipc: A61K 39/35 20060101ALI20120817BHEP

Ipc: A61K 39/145 20060101ALI20120817BHEP

Ipc: A61K 39/39 20060101ALI20120817BHEP

Ipc: A61K 39/385 20060101ALI20120817BHEP

Ipc: A61P 37/08 20060101ALI20120817BHEP

Ipc: A61K 38/17 20060101AFI20120817BHEP

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): DE FR GB IT

REG Reference to a national code

Ref country code: GB

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: DE

Ref legal event code: R096

Ref document number: 602006036063

Country of ref document: DE

Effective date: 20130627

RAP2 Party data changed (patent owner data changed or rights of a patent transferred)

Owner name: APPLIED MEDICAL ENZYME RESEARCH INSTITUTE CORPORAT

Owner name: THE UNIVERSITY OF TOKUSHIMA

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

REG Reference to a national code

Ref country code: DE

Ref legal event code: R082

Ref document number: 602006036063

Country of ref document: DE

Representative=s name: DEHNS, GB

Effective date: 20140220

Ref country code: DE

Ref legal event code: R081

Ref document number: 602006036063

Country of ref document: DE

Owner name: APPLIED MEDICAL ENZYME RESEARCH INSTITUTE CORP, JP

Free format text: FORMER OWNER: THE UNIVERSITY OF TOKUSHIMA, TOKUSHIMA, JP

Effective date: 20140220

26N No opposition filed

Effective date: 20140204

REG Reference to a national code

Ref country code: DE

Ref legal event code: R097

Ref document number: 602006036063

Country of ref document: DE

Effective date: 20140204

REG Reference to a national code

Ref country code: FR

Ref legal event code: PLFP

Year of fee payment: 11

REG Reference to a national code

Ref country code: FR

Ref legal event code: PLFP

Year of fee payment: 12

REG Reference to a national code

Ref country code: FR

Ref legal event code: PLFP

Year of fee payment: 13

REG Reference to a national code

Ref country code: DE

Ref legal event code: R082

Ref document number: 602006036063

Country of ref document: DE

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20210820

Year of fee payment: 16

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: IT

Payment date: 20220825

Year of fee payment: 17

Ref country code: DE

Payment date: 20220620

Year of fee payment: 17

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 20220823

Year of fee payment: 17

GBPC Gb: european patent ceased through non-payment of renewal fee

Effective date: 20220804

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: GB

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20220804

REG Reference to a national code

Ref country code: DE

Ref legal event code: R119

Ref document number: 602006036063

Country of ref document: DE

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20230804

Ref country code: FR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20230831

Ref country code: DE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20240301